MDA-MB-231, MDA-MB-468, SKBR3 and MCF-10A cells were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China). MDA-MB-231 cells were incubated in DMEM culture medium (Gibco, 11,965,092) with 10% foetal bovine serum (FBS). MDA-MB-468 cells were incubated in RPMI 1640 culture medium (Gibco, 11,875,093) with 10% FBS. SKBR3 cells were cultured in special culture medium (Procell, CM-0211) containing McCoy's 5A, 10% FBS and 1% penicillin/streptomycin. MCF-10A was cultured in special culture medium (Procell, CM-0525) containing DMEM/F12, 5% Horse serum, 20 ng/mL EGF, 0.5 μg/mL Hydrocortisone, 10 μg/mL Insulin, 1% NEAA and 1% P/S. Total RNA was isolated with the TRIzol reagent (Invitrogen). Complementary DNA was synthesized from 1 μg total RNA using MightyScript Plus First Strand cDNA Synthesis Master Mix (Sangon Biotech). Quantitative real-time polymerase chain reaction (qRT-PCR) was performed with a PowerUp SYBR Green Master Mix (Thermo Fisher, A25742) after RNA extraction and reverse transcription from all four cell lines. Relative mRNA levels were calculated using the comparative Ct method (ΔCt). The primer sequences are listed in Supplementary Material S1 .
Skbr3
Manufactured by Procell
Sourced in China
SKBR3 is a cell line derived from a human breast adenocarcinoma. It is a well-characterized model system widely used in cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Mda mb 231, by Procell (14 mentions)
Mcf 10a, by Procell (12 mentions)
Mcf 7, by Procell (12 mentions)
Mda mb 468, by Procell (5 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Mcf 7, by Thermo Fisher Scientific (5 mentions)
Bt 549, by Procell (4 mentions)
Mda mb 231, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Mda mb 468, by Thermo Fisher Scientific (2 mentions)
Cm 0211, by Procell (2 mentions)
Zr 75 1, by Procell (2 mentions)
Skbr3, by American Type Culture Collection (2 mentions)
Mcf 10a, by American Type Culture Collection (2 mentions)
Mda mb 231, by American Type Culture Collection (2 mentions)
Dmem f12, by Procell (2 mentions)
Bt 474, by Procell (2 mentions)
Hcc1937, by Procell (2 mentions)
Sk br 3, by Thermo Fisher Scientific (2 mentions)
21 protocols using skbr3
1
Gene Expression Analysis of Breast Cancer Cell Lines
2
Cultivation of Breast Cancer Cell Lines
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Human breast cancer cell lines ZR-75-1, MCF7, JIMT1, MDA-MB-468, MDA-MB-231 and SKBR3 were purchased from Procell (https://www.procell.com.cn
3
Culturing Human Breast Cell Lines
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The human breast epithelial cell line MCF-10A and cancer cell lines (MCF-7, MDA-MB-231, MDA-MB-468, SK-BR-3 and HCC70) were purchased from Procell Life Science&Technology Co., Ltd. MCF-10A was grown in MCF-10A cell special medium (CM-0525, Procell, Wuhan, China). Those breast cancer cells were grown in DMEM medium (Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum, 100 ug/mL streptomycin and 100 U/mL penicillin at 37°C with an atmosphere of 5% CO2 and 95% humidity.
4
Culturing Breast Cancer Cell Lines
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SK-BR-3 (human HER2-positive breast cancer cell line), MCF-10A (normal human breast cell line), and MDA-MB-231 (triple-negative breast cancer cell line) were obtained from the American Type Culture Collection (ATCC; Rockville, MD, United States). SK-BR-3 cell was cultured in DMEM medium supplemented with 10% FBS and 1% penicillin–streptomycin (100 IU/ml), MCF-10A cell was cultured in MCF-10A special medium (Procell Life Science and Technology, China). MDA-MB-231 cell was cultured in DMEM/F12 medium supplemented with 10% FBS and 1% penicillin–streptomycin (100 IU/ml). All the cells were incubated at 37°C in a 5% CO2 humidified incubator.
5
Breast Cancer Cell Line Characterization
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The human breast cancer cell lines (T47D, BT549, MCF7, MDA-MB-468, MDA-MB-231, and SKBR3) were purchased from Shaanxi Yike Biotechnology (Xi’an, China). MCF10A cells were cultured in DMEM/F12 (Procell) supplemented with 5% horse serum and 10 μg/mL insulin (Sigma). SKBR3 cells were grown in McCoy’s 5A medium (Procell) containing 10% fetal bovine serum (FBS). T47D and BT549 cells were cultured in a RPMI-1640 medium (Gibco) supplemented with 10% FBS. The MCF7, MDA-MB-231, and MDA-MB-468 cells were grown in DMEM (Gibco) supplemented with 10% FBS. All cells were incubated in a 5% CO2 incubator at 37 °C. Dasatinib (HY-10181) and Adezmapimod (HY-10256) were purchased from MedChemExpress (MCE, Shanghai, China). Phorbol 12-myristate 13-acetate (PMA) was purchased from Solabio (P6741).
6
Breast Cancer Cell Line Culture
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Breast cancer cell lines (SK-BR-3 (HTB-30) and MCF7 (HTB-22)) and culture media were obtained from American Type Culture Collection (ATCC, USA). SK-BR-3 cells were cultivated in McCoy’s 5a Medium Modified (30–2007) supplemented with 10% fetal bovine serum (FBS, 164,210–500, Procell). MCF7 cells were cultured in Eagle’s Minimum Essential Medium (30–2003) added with 0.01 mg/ml human recombinant insulin (91077C, Sigma-Aldrich, USA) and 10% FBS. The incubation was carried out at 37°C with 5% CO2 (Heracell Vios 160i CR CO2 incubator, 51,033,770, Thermo Scientific, USA).
7
Overexpression of DNAJB4 in Breast Cancer Cells
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Human breast cancer cell lines (MCF-7; BT-549; SKBR3; MDA-MB-231) and normal breast epithelial cell line (MCF-10A) were purchased from Procell (Wuhan, China). Cells were cultured in in DMEM containing 10% fetal bovine serum (FBS; C0400, VivaCell, Shanghai, China) and 1% penicillin/streptomycin (C0222, Beyotime, China) at 5% CO2 and 37 °C. The DNAJB4 overexpression lentivirus and negative control were purchased from GeneChem (Shanghai, China). The DNAJB4 overexpression lentivirus and negative control virus (MOI = 40) were transfected into MCF-7 cells using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer's protocol.
8
MTT Cytotoxicity Assay Protocol
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MTT assays were performed as previously described.11,12 (link) Briefly, cells were seeded into 96-well plates at a density of 5 × 103 cells per well for 24 h, and were the exposed to different concentrations of test compounds. After incubation for 72 h, cells were stained with 25 μL of MTT solution (5 mg mL−1) for 25 min. Finally, the mixture of medium and MTT solution was removed and 75 μL DMSO was added to dissolve formazan crystals. Absorbance of each well was measured at 544 nm (test wavelength) and 690 nm (background) using the Multi-Mode Microplate Reader. Background was subtracted from the absorbance of each well. A549 (human lung cancer) and Huh7 (human hepatoma), and HepG2 (human hepatoma) was purchased from the American Type Culture Collection (ATCC). SK-BR-3 (human breast cancer) was purchased from the Procell Life Science&Technology Co., Ltd J82 (human bladder cancer cell line) and MB49 (WT) (mouse bladder carcinoma cell line) was obtained from the Merck Millipore. MB49 (STR), the cisplatin resistant subclone of MB49 cells, was established in our lab by weekly exposure to cisplatin.
9
Culturing Human Breast Cell Lines
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Normal human breast epithelial cells (MCF-10A) and human breast cancer cells (MDA-MB-231, HCC1937, MCF-7, SK-BR-3 and BT474) were provided from Procell Life Science & Technology Co., Ltd. The MCF-10A cells were cultured in Dulbecco's modified Eagle's medium (DMEM)/F12 (Procell Life Science & Technology Co., Ltd.) containing 5% horse serum (Procell Life Science & Technology Co., Ltd.), 20 ng/ml EGF, 0.5 µg/ml hydrocortisone, 10 µg/ml insulin, 1% non-essential amino acids and 1% penicillin/streptomycin. The breast cancer cells were cultured in DMEM (Hyclone; Cytiva) containing 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific; Inc.) and 1% penicillin/streptomycin. All cells were cultured at 37˚C in a humidified atmosphere of 95% air and 5% CO2.
10
Evaluating EZR Knockdown in BC Cell Lines
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BC cell lines (MCF-7, MCF-7/ADR, BT-549, BT-549/ADR, MDA-MB-468 and SKBR-3) were purchased from Procell Company (Wuhan, China). MCF-7 cells were cultured in DMEM (Gibco, USA) and BT-549 cells were RPMI 1640 (HyClone, USA). MCF-7/ADR cells were grown in DMEM containing 10% FBS and1 µg/mL ADR. BT-549/ADR cells were grown in RPMI 1640 containing 10% FBS and1 µg/mL ADR. MDA-MB-468 cells were cultured in DMEM/F12 medium with 10% FBS. SKBR-3cells were plated in DMEM supplemented with 10% FBS. All media were supplemented with 10% fetal bovine serum (BI, Israel) and 1% penicillin/streptomycin (HyClone, USA). The cells were grown in a 37°C humidified incubator with 5% CO2. EZR siRNAs (si-EZR), scrambled negative control (NC) siRNAs (si-NC), empty pcDNA3.0 vector and EZR-pcDNA3.0 vector were synthesized by Gene Pharma (Shanghai, China), as follows: EZR 5′-UUCUCAUAAAUAUUCAGUCCAAGGG-3′, 5′-UUCUGCGTACCUAUCACGUTT-5′. Briefly, cells were plated in six-well plates at a density of 5 × 105 cells/well overnight and then transfected with 50 nm si-EZR or si-NC by using Lipofectamine 3000 reagent (Invitrogen, USA).
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